title: Identification of species belonging to the Bifidobacterium genus by PCR-RFLP analysis of a hsp60 gene fragment.
abstract: BACKGROUND
  Bifidobacterium represents one of the largest genus within the Actinobacteria, and includes at present 32 species. These species share a high sequence homology of 16S rDNA and several molecular techniques already applied to discriminate among them give ambiguous results.
  
  
  RESULTS
  This work illustrates a simple and cheap molecular tool for the identification of Bifidobacterium species. The hsp60 universal primers were used in a simple PCR procedure for the direct amplification of 590 bp of the hsp60 sequence. The in silico restriction analysis of bifidobacterial hsp60 partial sequences allowed the identification of a single endonuclease (HaeIII) able to provide different PCR-restriction fragment length polymorphism (RFLP) patterns in the Bifidobacterium spp. type strains evaluated. The electrophoretic analyses allowed to confirm the different RFLP patterns.
  
  
  CONCLUSIONS
  The developed PCR-RFLP technique resulted in efficient discrimination of the tested species and subspecies and allowed the construction of a dichotomous key in order to differentiate the most widely distributed Bifidobacterium species as well as the subspecies belonging to B. pseudolongum and B. animalis.
authors.forename: Loredana
authors.initials: L
authors.lastname: Baffoni
authors.forename: Verena
authors.initials: V
authors.lastname: Stenico
authors.forename: Erwin
authors.initials: E
authors.lastname: Strahsburger
authors.forename: Francesca
authors.initials: F
authors.lastname: Gaggìa
authors.forename: Diana
authors.initials: D
authors.lastname: Di Gioia
authors.forename: Monica
authors.initials: M
authors.lastname: Modesto
authors.forename: Paola
authors.initials: P
authors.lastname: Mattarelli
authors.forename: Bruno
authors.initials: B
authors.lastname: Biavati
mesh_terms: D001644:Bifidobacterium / Q000145:classification* / Q000235:genetics; D018834:Chaperonin 60 / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA
chemical_list: D018834:Chaperonin 60; D004269:DNA, Bacterial